Background: Thuja articulata is a Mediterranean forest species from the Cupressaceae family, it has been used in popular medicine to treat several diseases. Various studies have been carried out in vitro using diverse T. articulata extracts to understand its traditional use. Methods: In this study, the ethyl acetate extract of T. articulata trunk barks was chemically identified using HPLC-PDA-ESI-MS, then examined in vitro for its antioxidant and α-amylase inhibitory effects. A molecular docking study was also performed to reinforce the noted bioactivities. Results: HPLC-PDA-ESI-MS analysis led to the identification of 22 polyphenolic compounds in the ethyl acetate extract of T. articulata trunk barks. This extract revealed interesting in vitro antioxidant properties and a significant α-amylase inhibitory action (IC50 = 16.08 ± 1.27 μg/mL). In silico analysis was found to agree with the in vitro studies in which major constituents of the ethyl acetate extract revealed low binding energy and a correct mode of interaction in the active pocket of the enzyme (PDB: 7TAA). The anti-a-amylase potential could be due either to a synergistic or individual action of certain constituents present in this extract. Conclusion: This study illustrates that the ethyl acetate extract of T. articulata trunk barks has potent sources of antioxidants and α-amylase inhibitors to be explored.